lncRNA OIP5-AS1 knockdown or miR-223 overexpression can alleviate LPS-induced ALI/ARDS by interfering with miR-223/NLRP3-mediated pyroptosis
lncRNA OIP5-AS1 knockdown or miR-223 overexpression can alleviate LPS-induced ALI/ARDS by interfering with miR-223/NLRP3-mediated pyroptosis
复制标题
LncRNA OIP5-AS1 敲低或 miR-223 过表达可以通过干扰 miR-223/NLRP3 介导的焦亡来减轻 LPS 诱导的 ALI/ARDS
DOI：
10.1002/jgm.3385
复制
发表时间：
2022-02-09
影响因子：
3.5
通讯作者：
Sun, Gengyun
中科院分区：
文献类型：
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作者：
Ji, Juanjuan;Ye, Wei;Sun, Gengyun
背景急性肺损伤（ALI）和急性呼吸窘迫综合征（ARDS）是危及生命的疾病，内皮屏障损伤是ALI/ARDS发病的重要因素。长链非编码 (lncRNA) 已被证明参与 ALI/ARDS 的进展。本研究旨在探讨 lncRNA opa 相互作用蛋白 5 反义 RNA 1 (OIP5-AS1) 在脂多糖 (LPS) 诱导的 ALI/ARDS 中的功能。方法采用聚合酶链式反应检测ALI/ARDS患者或健康供者血清中OIP5-AS1和miR-223的水平。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑溴化物测定法检测人肺微血管内皮细胞 (HPMEC) 的增殖情况。流式细胞术检测HPMECs的凋亡情况。通过Western blotting检测NLRP3、ASC、GSDMD-N和caspase-1的蛋白水平来检测HPMECs的焦亡情况。采用酶联免疫吸附法检测白细胞介素（IL）-1β、IL-6、IL-18和IL-10，以测定HPMEC的炎症反应。测量活性氧、超氧化物歧化酶和丙二醛的产生，以确定 HPMEC 的氧化应激。 OIP5-AS1 和 miR-223 的靶标由 StarBase 预测，并通过双荧光素酶报告基因测定进行确认。结果我们发现 ALI/ARDS 患者和 LPS 处理的 HPMEC 血清中 OIP5-AS1 上调，miR-223 下调。从功能上来说，敲除 OIP5-AS1 可诱导 LPS 处理的 HPMEC 增殖并抑制细胞凋亡、细胞焦亡、炎症反应和氧化应激。有趣的是，miR-223 是 OIP5-AS1 的靶标，抑制 miR-223 消除了 si-OIP5-AS1 对 LPS 诱导的 HPMEC 的影响。更重要的是，miR-223直接靶向NLRP3，miR-223过表达可促进LPS处理的HPMEC的增殖并抑制细胞凋亡、细胞焦亡、炎症反应和氧化应激，而NLRP3过表达可消除这种作用。最后，我们发现 OIP5-AS1 敲低和 miR-223 过表达都可以减轻体内 LPS 诱导的 ALI/ARDS。结论 综上所述，我们发现lncRNA OIP5-AS1通过miR-223/NLRP3轴加重LPS诱导的ALI/ARDS，为ALI/ARDS治疗提供新靶点。
Background Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are life-threatening diseases and endothelial barrier injury is an important contributor to the pathogenesis of ALI/ARDS. Long non-coding (lncRNA) has been shown to participate in the progression of ALI/ARDS. The present study aimed to investigate the function of lncRNA opa-interacting protein 5 antisense RNA 1 (OIP5-AS1) in lipopolysaccharide (LPS)-induced ALI/ARDS. Methods OIP5-AS1 and miR-223 levels were detected by a polymerase chain reaction in the serum of ALI/ARDS patients or healthy donors. An 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay was performed to detect the proliferation of human pulmonary microvascular endothelial cells (HPMECs). Flow cytometry were performed to detect the apoptosis of HPMECs. The protein levels of NLRP3, ASC, GSDMD-N and caspase-1 were measured by western blotting to detect the pyroptosis of HPMECs. Interleukin (IL)-1 beta, IL-6, IL-18 and IL-10 were detected by an enzyme-linked immunosorbent assay to measure the inflammatory response of HPMECs. The production of reactive oxygen species, superoxide dismutase and malondialdehyde was measured to determine the oxidative stress of HPMECs. Targets of OIP5-AS1 and miR-223 were predicted by StarBase and confirmed by a dual-luciferase reporter assay. Results We found that OIP5-AS1 was up-regulated and miR-223 was down-regulated in the serum of ALI/ARDS patients and LPS-treated HPMECs. Functionally, knockdown of OIP5-AS1 induced proliferation and inhibited apoptosis, pyroptosis, inflammatory response and oxidative stress of LPS-treated HPMECs. Interestingly, miR-223 was a target of OIP5-AS1 and miR-223 inhibition abolished the effects of si-OIP5-AS1 on LPS-induced HPMECs. More importantly, miR-223 directly targeted NLRP3, and miR-223 overexpression promoted proliferation and inhibited apoptosis, pyroptosis, inflammatory response and oxidative stress of LPS-treated HPMECs, with this being abolished by NLRP3 overexpression. Finally, we found that OIP5-AS1 knockdown and miR-223 overexpression could both alleviate LPS-induced ALI/ARDS in vivo. Conclusions Taken together, we find that lncRNA OIP5-AS1 aggravates LPS-induced ALI/ARDS via miR-223/NLRP3 axis and provides new targets for ALI/ARDS therapy.